HPLC-MS/MS测定人脑脊液中多黏菌素E1和E2的方法及临床应用  

Determination of Polymyxin E1 and E2 in Human Cerebrospinal Fluid by High Performance Liquid Chromatography-Tandem Mass Spectrometry

作　　者：王琴会 张婷婷 张文娟 强琼 任静 WANG Qin-hui;ZHANG Ting-ting;ZHANG Wen-juan;QIANG Qiong;Ren Jing(The Second Affiliated Hospital of Air Force Medical University,Xi'an 710038,China)

机构地区：[1]空军军医大学第二附属医院,陕西西安710038

出　　处：《解放军药学学报》2024年第4期308-313,共6页Pharmaceutical Journal of Chinese People's Liberation Army

摘　　要：目的建立一种新的盐析辅助液-液萃取结合高效液相色谱-串联质谱联用技术,快速测定人脑脊液中多黏菌素E1和E2浓度的方法。方法以多黏菌素B1为内标,用乙腈萃取脑脊液样本,甲酸铵作为盐析剂进行液-液萃取。并对盐析剂浓度、盐析剂体积以及萃取剂体积进行了优化。色谱柱采用Waters Xselect■HSS T3(2.1 mm×150 mm,2.5μm),流动相用乙腈和0.1%甲酸水溶液,采用梯度洗脱方式,检测时间6 min。结果在优选实验条件下,脑脊液中多黏菌素E1和E2分别在0.20~8.00μg·ml^(-1)(r=0.9960),0.35~14.12μg·ml^(-1)(r=0.9984)范围线性良好。待测物的准确度均在91.4%~109.0%范围,日内、日间精密度的相对标准偏差均低于9.5%。多黏菌素E1的提取回收率为77.7%~84.3%,多黏菌素E2的提取回收率为60.3%~68.3%。待测物在脑脊液中无明显基质效应。脑脊液样品在采集、储存、预处理过程中均能保持稳定。结论该方法操作简便,实验成本低,准确度高,重现性好,可应用于临床患者脑脊液中多黏菌素E1和E2的浓度测定。Objective To establish a new salting-out-assisted liquid-liquid extraction combined with high performance liquid chromatography tandem mass spectrometry(HPLC-MS/MS)technique for the rapid determination of polymyxin E1 and E2 concentrations in human cerebrospinal fluid.Methods Polymyxin B1 was used as an internal standard,the cerebrospinal fluid samples were extracted with acetonitrile,and ammonium formate was used as the salting-out agent for liquid-liquid extraction.The concentration and volume of the salting-out agent as well as the volume of the extracting agent were optimized.A Waters XselectHSS T3 column(2.1 mm×150 mm,2.5μm)was used.The mobile phase consisted of acetonitrile and water containing 0.1%formic acid.The detection length was 6 min by gradient elution.Results Under the optimal experimental conditions,the calibration curves of analytes in cerebrospinal fluid showed good linearity within the range of 0.20-8.00μg·ml^(-1)for polymyxin E1(r=0.9960)and 0.36-14.4μg·ml^(-1)for polymyxin E2(r=0.9984).The accuracies of the analytes ranged from 91.4%to 109.0%,and the relative standard deviations(RSD)of intra-and inter-day precisions were less than 9.5%.The recoveries of polymyxin E1 and polymyxin E2 were 77.7%-84.3%and 60.3%-68.3%respectively.The analytes had no obvious matrix effect in cerebrospinal fluid.The cerebrospinal fluid samples were stable during collection,storage and pretreatment.Conclusion This method is simple,inexpensive,highly accurate and reproducible,which can be applied to determination of polymyxin E1 and E2 in human cerebrospinal fluid.

关 键 词：高效液相色谱-串联质谱联用 盐析辅助液-液萃取 脑脊液 多黏菌素E 

分 类 号：R917[医药卫生—药物分析学]